I have spent the last ten years building an expertise in the study of cAMP microdomains, specifically those regulated by the novel protein soluble adenylyl cyclase (sAC). During my training in dermatology, I applied the techniques and skills I have developed to the understanding of hyper proliferative keratinocyte skin disease, which led to a recent publication in the Journal of Investigative Dermatology. Over the past year, I began to explore the role of cAMP signaling in melanoma. I have developed two fruitful collaborations with experts in melanoma biology, Dr. Wolchok and Dr. Bastian at Memorial Sloan Kettering Cancer Center (MSKCC), and have continued my collaboration with an expert in sAC biology, Dr. Buck. Dr. Bastian and Dr. Buck will serve as my mentors during my training, and Dr. Wolchok will serve as an advisor. Over the next five years, I will have the privilege of being trained at both a world-class cancer research institution, MSKCC, and a leading medical college, Weill Cornell Medical Center (WCMC), both located at one of the most prestigious research corners in the world (WCMC/MSKCC/Rockefeller), proximity to which allows for unprecedented resources and exposure. Over the next five years, I have formulated a plan to further my expertise in cAMP signaling and develop my knowledge in melanoma biology. My short- term plan is to continue my education in order to become a physician scientist in dermatology. My long-term goal, in five years, is to secure R01 funding as an independent investigator. Melanoma is the sixth most common human cancer and one of the deadliest. Once melanoma spreads beyond the skin, effective medical interventions are limited. While certain signaling pathways, such as the Ras/Raf/ERK pathway, have well- established roles in controlling melanoma pathogenesis, recent clinical trials and basic science reports suggest that blocking this pathway alone is an insufficient treatment for melanoma. It has become apparent that the development of an effective treatment for melanoma relies on an improved understanding of the intricacies of melanoma signaling. cAMP has been implicated in a variety of signaling pathways that are key to the development and pathogenesis of melanoma, but in many cases the source and ramifications of cAMP signaling is poorly understood. A new class of adenylyl cyclases called soluble adenylyl cyclase (sAC) was recently characterized, and I was involved in developing a cadre of novel small molecule and genetic inhibitors capable of differentiating sAC-generated and the canonical G-protein sensitive transmembrane adenylyl cyclase (tmAC)-generated cAMP. In the past, I have used these reagents to establish the role of sAC in nerve growth factor-induced Rap activation in PC-12 cells, netrin-induced growth cone chemotaxis in primary neurons, and glucose-induced cAMP in beta cells. In each of these cases, involvement of cAMP was known, but a suitable model to explain its role was lacking. By differentiating tmAC- from sAC-dependent cAMP, more coherent models have been developed. I have applied these techniques to melanoma biology and have found that sAC protein migrates into the nucleus in human melanocytic proliferations as melanocytes acquire cellular atypical. In the past, I demonstrated that nuclear sAC activates the cAMP responsive binding protein transcription factor. I have also discovered that sAC protein is upregulated in melanoma, and inhibitors of sAC block melanocyte growth. In this proposal I plan to evaluate the relative contributions of sAC and tmAC microdomains in melanoma pathogenesis. I predict that this line of investigation will lead to novel therapeutic targets of melanoma.! PUBLIC HEALTH RELEVANCE: Melanoma is a deadly cancer leading to over 8,000 deaths a year. cAMP signaling pathways are implicated in melanoma pathogenesis but the mechanisms are not fully understood. I have developed a novel method for studying cAMP signaling and propose to use this method to better understand melanoma pathogenesis. The written critiques and criteria scores of individual reviewers are provided in essentially unedited form in the "Critique" section below. Please note that these critiques and criteria scores were prepared prior to the meeting and may not have been revised subsequent to any discussions at the review meeting. The "Resume and Summary of Discussion" section above summarizes the final opinions of the committee.